Imbalances in the essential mineral elements copper, magnesium and zinc have been demonstrated in a number of immunologically related diseases. These three elements have been reported to vary from normal balances in autoimmune diseases (i.e., Crohn's diseaae rheumatoid arthritis, systemic lupus erythematosus, (etc.), cancer and infectious diseases, but little is known on how these tlements effect the immune response. Several groups have reported on the role of zinc in T cell functions and specifically the effects of zinc deficiency on T-helper cell activity in long-term in vivo dietary zinc deficiency models. Problems in interpreting these data have developed since immune cells were cultured in zinc containing media when the assays for immunological reactivity were done. We have developed an in vitro culture method that permits us to control the elemental environment of the immune cell populations and to study the generation of immune responses under in vitro elemental deficiency conditions. This proposal is developed to study: at what step(s) in the generation of cytotoxic T lymphocytes (CTLs) the essential elements copper, magnesium and zinc are involved, to specifically define mechanisms dependent on the three minerals. To accomplish this aim, we will use in vitro allo-antigen stimulated mixed lynphocyte cultures as we first study the involvement of copper, magnesium and zinc in three phases of CTL generation: the initiation phase, proliferation and the lytic phase. Studies will focus on determining at what step and at what level of mineral deficiency CTL generation is modified by the lack of metals. Second, we will compare the changes in CTL generation of splenocytes from mice dietarily made deficient in copper, magnesium or zinc with splenocytes that have been made deficient by in vitro methods. These comparisons are necessary to ascertain the impact metal deficiencies and what role in vitro culture has on the reactivity of T cells. Thire, we will study the effects of deficiencies of copper, magnesium and zinc on the "helper" factors, Interleukin-1 (IL-1) and Interleukin-2 (IL-2), both of which are required for CTL generation. Using in vitro methods, we will study the synthesis and release of IL-1 and IL-2 and receptors for IL-2 in response to single element deficiencies (copper, magnesium or zinc).